Effects of prenatal dexamethasone exposure on adult C57BL/6J mouse metabolism and oxidative stress.

Prenatal glucocorticoid exposure has been shown to alter hypothalamic-pituitary-adrenal axis function resulting in altered fetal development that can persist through adulthood. Fetal exposure to excess dexamethasone, a synthetic glucocorticoid, has been shown to alter adult behaviour and metabolism. This study investigated the effects prenatal dexamethasone exposure had on adult offspring cardiac and liver metabolism and oxidative stress. Pregnant C57BL/6 mice received a dose of 0.4 mg/kg dexamethasone on gestational days 15-17. Once pups were approximately 7 months old, glucose uptake was determined using positron emission tomography and insulin resistance (IR) was determined by homeostatic model assessment (HOMA) IR calculation. Oxidative stress was assessed by measuring 4-hydroxynonenal protein adduct formation and total reactive oxygen species. Female dexamethasone group had significantly increased glucose uptake when insulin stimulated compared to vehicle-treated mice. HOMA IR revealed no evidence of IR in either male or female offspring. There was also no change in oxidative stress markers in either cardiac or liver tissues of male or female offspring. These data suggest that prenatal dexamethasone exposure in male mice does not alter oxidative stress or metabolism. However, prenatal dexamethasone exposure increased glucocorticoids, cardiac glucose uptake, and pAkt signaling in female heart tissues in adult mice, suggesting there are sex differences in prenatal dexamethasone exposure.

The Role of DNMT and HDACs in the Fetal Programming of Hypertension by Glucocorticoids.

The causes of hypertension are complex and involve both genetic and environmental factors. Environment changes during fetal development have been linked to adult diseases including hypertension. Studies show that timed in utero exposure to the synthetic glucocorticoid (GC) dexamethasone (Dex) results in the development of hypertension in adult rats. Evidence suggests that in utero stress can alter patterns of gene expression, possibly a result of alterations in the topology of the genome by epigenetic markers such as DNA methyltransferases (DNMTs) and histone deacetylases (HDACs). The objective of this study was to determine the effects of epigenetic regulators in the fetal programming and the development of adult hypertension. Specifically, this research examined the effects of the HDAC inhibitor valproic acid (VPA) and the DNMT inhibitor 5-aza-2'-deoxycytidine (5aza2DC) on blood pressure (BP) and gene expression in prenatal Dex-programmed rats. Data suggest that both VPA and 5aza2DC attenuated the Dex-mediated development of hypertension and restored BP to control levels. Epigenetic DNMT inhibition (DNMTi) or HDAC inhibition (HDACi) also successfully attenuated elevations in the majority of altered catecholamine (CA) enzyme expression, phenylethanolamine N-methyltransferase (PNMT) protein, and elevated epinephrine (Epi) levels in males. Although females responded to HDACi similar to males, DNMTi drove increased glucocorticoid receptor (GR) and PNMT expression and elevations in circulating Epi in females despite showing normotensive BP.

Reactive oxygen and nitrogen species balance in the determination of thyroid hormones-induced cardiac hypertrophy mediated by renin-angiotensin system.

Role of reactive oxygen species (ROS)/nitric oxide (NO) balance and renin-angiotensin system in mediating cardiac hypertrophy in hyperthyroidism was evaluated in an in vivo and in vitro experimental model. Male Wistar rats were divided into four groups: control, thyroid hormone, vitamin E (or Trolox, its hydrosoluble analogue), thyroid hormone+vitamin E. Angiotensin II receptor (AT1/AT2) gene expression, immunocontent of AT1/AT2 receptors, angiotensinogen, NADPH oxidase (Nox2), and nitric oxide synthase isoforms, as well as ROS concentration (hydrogen peroxide and superoxide anion) were quantified in myocardium. Thyroid hormone increased ROS and NO metabolites, iNOS, nNOS and eNOS isoforms and it was accompanied by cardiac hypertrophy. AT1/AT2 expression and the immunocontent of angiotensinogen and Nox2 were enhanced by thyroid hormone. Antioxidants reduced ROS levels, Nox2, AT1/AT2, NOS isoforms and cardiac hypertrophy. In conclusion, ROS/NO balance may play a role in the control of thyroid hormone-induced cardiac hypertrophy mediated by renin-angiotensin system.

The role of redox signaling in cardiac hypertrophy induced by experimental hyperthyroidism.

This study was conducted to test whether oxidative stress activates the intracellular protein kinase B (AKT1) signaling pathway, which culminates with cardiac hypertrophy in experimental hyperthyroidism. Male Wistar rats were divided into four groups: control, vitamin E, thyroxine (T(4)), and T(4)+vitamin E. Hyperthyroidism was induced by T(4) administration (12 mg/l in drinking water for 28 days). Vitamin E treatment was given during the same period via s.c. injections (20 mg/kg per day). Morphometric and hemodynamic parameters were evaluated at the end of the 4-week treatment period. Protein oxidation, redox state (reduced glutathione, GSH/glutathione dissulfide, GSSG), vitamin C, total radical-trapping antioxidant potential (TRAP), hydrogen peroxide (H2O2), and nitric oxide metabolites (NO(X)) were measured in heart homogenates. The p-AKT1/AKT1 ratio, p-glycogen-synthase kinase (GSK)3B/GSK3B ratio, FOS, and JUN myocardial protein expression were also quantified by western blot after 4 weeks. Increases in biochemical parameters, such as protein oxidation (41%), H2O2 (62%), and NO(X) (218%), and increase in the left ventricular end-diastolic pressure were observed in the T(4) group. T(4) treatment also caused a decrease in GSH/GSSG ratio (83%), vitamin C (34%), and TRAP (55%). These alterations were attenuated by vitamin E administration to the hyperthyroid rats. Expression of p-AKT1/AKT1, p-GSK3B/GSK3B, FOS, and JUN were elevated in the T(4) group (by 69, 37, 130, and 33% respectively), whereas vitamin E administration promoted a significant reduction in their expression. These results indicate that oxidative stress plays an important role in cardiac hypertrophy, and suggest redox activation of AKT1 and JUN/FOS signaling pathways with H2O2 acting as a possible intracellular mediator in this adaptive response to experimental hyperthyroidism.

Activation of apoptotic processes during transition from hypertrophy to heart failure in guinea pigs.

Changes in oxidative stress and apoptotic process were studied during the progression of a compensated hypertrophy to a decompensated heart failure in guinea pigs. Banding of the ascending aorta resulted in heart hypertrophy. At 10 wk, ventricle-to-body weight ratio and thickness of the interventricular septum as well as the left ventricular wall were increased significantly. Although fractional shortening and ejection fraction were decreased, there were no signs of heart failure. Furthermore, there was no increase in wet-to-dry weight ratios for the lungs and liver at this stage. However, at 20 wk, heart failure was characterized by a significant depression in heart function as indicated by a decrease in fractional shortening, and ejection fraction and a lesser increase in wall thickness from diastole to systole. Animals also showed clinical signs of heart failure, and the wet-to-dry weight ratios of the lungs and liver were significantly higher. Cardiomyocyte oxidative stress was significantly higher in the 20-wk aortic-banded group. The ratio of Bax to Bcl-xl showed an increase at 10 wk, and there was a further increase at 20 wk. Mitochondrial membrane potential in the aortic-banded animals was significantly decreased at 10 and 20 wk. Cytochrome c levels were higher in the cytosol compared with the mitochondria, leading to a considerable increase in the expression of p17 subunit of caspase-3. At 20 wk, both early and late stages of apoptosis were observed in isolated cardiomyocytes. It is suggested that an increase in oxidative stress initiates mitochondrial death pathway during the hypertrophic stage, leading to apoptosis and heart failure at a later stage.

Antioxidant enzyme gene expression in congestive heart failure following myocardial infarction.

Increased oxidative stress and reduction in antioxidant enzymes have been suggested to be involved in the pathophysiology of congestive heart failure subsequent to myocardial infarction (MI). The objective of the present study was to characterize changes in the mRNA abundance and protein levels for the enzymatic antioxidants, superoxide dismutase (SOD), glutathione peroxidase (GSHPx) and catalase during the sequelae of congestive heart failure in rats. MI was produced by the ligation of the left coronary artery and hearts from controls and 1, 4 and 16 week PMI groups were analyzed. Losartan treatment (2 mg/ml in drinking water, daily) was started at 4 weeks and continued for 12 weeks. The mRNA levels for SOD were reduced by about 40% at 1-week PMI, were near to the control levels at 4-week PMI and at 16 weeks PMI, the levels were reduced by about 73% below the controls. GSHPx mRNA levels remained unchanged at all time points. The mRNA levels for catalase remained unchanged at 1 and 4 weeks PMI and were significantly reduced by about 44% at 16 weeks PMI as compared to the controls. The protein levels for MnSOD, CuZnSOD, GSHPx at 1 and 16 weeks remained unchanged in treated and untreated PMI groups. However, the protein levels for catalase was significantly increased in the control and PMI groups treated with Losartan. It is concluded that changes in the SOD and catalase activities during severe heart failure correlated with changes in mRNA for these enzymes. The precise mechanism/s for the improvement in antioxidant reserve and protein levels after Losartan treatment is/are unclear at this time.

Modulation of oxidative stress by a selective inhibition of angiotensin II type 1 receptors in MI rats.

OBJECTIVES: To examine whether blocking of the renin-angiotensin system (RAS) at the angiotensin II type 1 (AT1) receptor site is accompanied by changes in the oxidative stress parameters. BACKGROUND: Congestive heart failure in rats after myocardial infarction (MI) has been shown to correlate with a decrease in antioxidant enzyme activities and an increase in oxidative stress. Inhibition of the RAS with captopril improves cardiac function and survival in MI rats with a reduction in oxidative stress. METHODS: Myocardial infarction in rats was produced by ligation of the left coronary artery. At four weeks after surgery, animals from the sham as well as MI groups were treated with losartan (2 mg/ml in drinking water daily). At 16 weeks after surgery, the animals were examined for hemodynamic function and the hearts were analyzed for antioxidant enzyme activities (superoxide dismutase, glutathione peroxidase and catalase) and oxidative stress (lipid hydroperoxides, reduced and oxidized glutathione and redox ratio). RESULTS: Congestive heart failure was characterized by dyspnea, depressed hemodynamic function and presence of lung and liver congestion. This was also associated with a decrease in the myocardial catalase (-25%), glutathione peroxidase (-38%) and superoxide dismutase (-42%) activities. An increase in oxidative stress in these hearts was indicated by an increase in lipid hydroperoxides (+67%) and reduction in the redox ratio (-75%). Hemodynamic function was better maintained and there were no indications of dyspnea or lung or liver congestion in the losartan-treated MI rats. In these animals, myocardial oxidative stress was markedly reduced and glutathione peroxidase and catalase activities were significantly improved compared with the untreated MI group. CONCLUSIONS: Blocking of RAS at the AT1 receptor site without the inhibition of angiotensin-converting enzymes modulates heart failure after MI, and this beneficial effect is associated with a decrease in oxidative stress. This study suggests a newer role for losartan in the treatment of heart failure.

Oxidative stress in congestive heart failure.

Heart failure is considered to be a complex clinical syndrome, with alterations in the multiple neurohumoral systems and subcellular cardiac sites that correlate with abnormal cardiac function. Strong evidence for the role of oxidative stress in the pathogenesis of heart failure has been provided by studies on experimental animals as well as humans. This concept is gaining more acceptance due to the fact that during heart failure, changes in different neurohormones, cytokines, nitric oxide, and activated inflammatory cells are closely linked to oxidative stress at the cellular and molecular levels. The present article provides a simple description of oxygen free radicals as well as the antioxidant defense system. Evidence for the role of oxidative stress in the pathogenesis of heart failure is reviewed in a concise manner.

Effects of methionine on endogenous antioxidants in the heart.

The deficiency of methionine, an essential amino acid, is associated with cardiovascular lesions. Because different types of cardiac pathologies are caused by a decrease in antioxidants, we examined the effects of methionine on myocardial antioxidant enzymes in hemodynamically assessed rats that were treated with methionine (10 mg/ml) in drinking water for 12, 24, and 48 h. Glutathione peroxidase (GSHPx) activity was significantly increased to 150.5 +/- 12.2 and 191.7 +/- 13.7% of the control value at 12 and 24 h, respectively, followed by a decline to 120 +/- 24.6% at 48 h. The mRNA levels of GSHPx at these time points were 151.2 +/- 12.0, 218.7 +/- 35.3, and 173.5 +/- 25.2%, respectively. Superoxide dismutase (SOD) activity was 144.3 +/- 3.7, 114.3 +/- 10.1, and 143.1 +/- 11. 2% at 12, 24, and 48 h, respectively. Catalase (Cat) activity was 272.4 +/- 5.4, 237.8 +/- 16.6, and 224.1 +/- 17.3% of the control value. The expression of Cat and SOD mRNA was unchanged at 12, 24, and 48 h. The lipid peroxidation was decreased by 24.4 +/- 11.2, 54. 9 +/- 0.1, and 6.4 +/- 2.1% at 12, 24, and 48 h, respectively. Methionine had no effect on the ventricular or aortic pressures, heart rate, and myocardial glutathione levels at any of the time points. The study shows that methionine has a significant effect on the myocardial antioxidant enzyme activities, and only changes in GSHPx enzyme activity correlated with the mRNA changes. These antioxidant changes may have a role in the beneficial effects of methionine in pathological rather than physiological conditions.

Metabolism of vitamin A in the heart increases after a myocardial infarction.

The supply of vitamin A to the myocardium by storage organs during increased oxidative stress subsequent to myocardial infarction (MI) was examined in hemodynamically assessed rats using compartment analysis of a radio-labeled vitamin A. 3H-Vitamin A was injected into two groups of rats: an MI group and a control group. There were no differences in the plasma or myocardial content of total vitamin A (unlabeled + labeled) between the two groups. However, the proportion of 3H-vitamin A was greater in the myocardium as well as plasma of MI rats. Rats with MI also had significantly lower 3H-vitamin A levels in liver and kidney than sham controls. The greatest difference in vitamin A content was in the concentrations of 3H-labeled storage forms of vitamin A in the liver of MI animals. Activity of bile salt-dependent retinyl ester hydrolase, an enzyme responsible for hydrolyzing vitamin A storage forms, was significantly increased in the liver of MI animals. These data indicate that analysis of plasma concentrations of vitamin A to ascertain links to cardiac conditions may be inappropriate. Specifically, during MI, increased amounts of vitamin A are mobilized from the liver to the heart without changing plasma concentrations. This is facilitated by an increase in the activity of an enzyme that hydrolyzes vitamin A storage forms.

Antioxidant potentials of vitamin A and carotenoids and their relevance to heart disease.

Despite being one of the first vitamins to be discovered, the full range of biological activities for vitamin A remains to be defined. Structurally similar to vitamin A, carotenoids are a group of nearly 600 compounds. Only about 50 of these have provitamin A activity. Recent evidence has shown vitamin A, carotenoids and provitamin A carotenoids can be effective antioxidants for inhibiting the development of heart disease. Vitamin A must be obtained from the diet: green and yellow vegetables, dairy products, fruits and organ meats are some of the richest sources. Within the body, vitamin A can be found as retinol, retinal and retinoic acid. Because all of these forms are toxic at high concentrations, they are bound to proteins in the extracellular fluids and inside cells. Vitamin A is stored primarily as long chain fatty esters and as provitamin carotenoids in the liver, kidney and adipose tissue. The antioxidant activity of vitamin A and carotenoids is conferred by the hydrophobic chain of polyene units that can quench singlet oxygen , neutralize thiyl radicals and combine with and stabilize peroxyl radicals. In general, the longer the polyene chain, the greater the peroxyl radical stabilizing ability. Because of their structures, vitamin A and carotenoids can autoxidize when O2 tension increases, and thus are most effective antioxidants at low oxygen tensions that are typical of physiological levels found in tissues. Overall, the epidemiological evidence suggests that vitamin A and carotenoids are important dietary factors for reducing the incidence of heart disease. Although there is considerable discrepancy in the results from studies in humans regarding this relationship, carefully controlled experimental studies continue to indicate that these compounds are effective for mitigating and defending against many forms of cardiovascular disease. More work, especially concerning the relevance of how tissue concentrations, rather than plasma levels, relate to the progression of tissue damage in heart disease is required. This review assembles information regarding the basic structure and metabolism of vitamin A and carotenoids as related to their antioxidant activities. Epidemiological, intervention trials and experimental evidence about the effectiveness of vitamin A and carotenoids for reducing cardiovascular disease is also reviewed.

Regional differences in non-enzymatic antioxidants in the heart under control and oxidative stress conditions.

Antioxidants are known to play an important role in mitigating oxidative stress injury. Regional concentrations of non-enzymatic antioxidants, redox ratio and lipid peroxides were studied in normal, ischemic and ischemic-reperfused rat hearts. Isolated perfused rat hearts were made globally ischemic for 45 min and reperfused for 15 min. Right ventricular wall (RVW), septum (S) and left ventricular wall (LVW) from control, ischemic (I) and reperfused (I-R) hearts were analysed. Tocopherol, retinol and ascorbic acid concentrations in different regions of perfused control hearts were not different. Reduced glutathione (GSH) was significantly lower in the RVW, while S and LVW had about three-fold higher levels. Oxidized glutathione (GSSG) was lower in the RVW and most concentrated in the LVW. The GSH:GSSG ratio was highest in the septum while RVW and LVW had similar values. Lipid hydroperoxide (LPx) concentrations in the three regions of control hearts were not different from each other. In I and I-R hearts, vitamin E declined in all three regions but the loss was significant only in the septum in the I group and in the septum and LVW of the I-R group. Vitamin A showed significant loss in all three regions of the I-R group. Vitamin C declined significantly only in the RVW of the I-R group. GSH increased in the RVW of the I and I-R groups compared to controls. GSSG was increased in the RVW and septum of the I group and in all regions of the I-R group. The redox ratio, GSH:GSSG, decreased in all regions of both I and I-R groups. LPx were increased in the septum of the I group and in all regions of the I-R group. Despite unique regional differences in non-enzymatic antioxidants, a comparable increase in LPx in the I-R group and similar extent of reduction in the redox ratio in different regions of the I and I-R groups, suggest that each myocardial region may use different antioxidant mechanisms to withstand oxidative stress.

Modulation of adriamycin-induced changes in serum free fatty acids, albumin and cardiac oxidative stress.

Adriamycin-induced cardiomyopathic changes are prevented by combination therapy with probucol. These beneficial effects are suggested to be due to a combination of antioxidant as well as lipid-lowering effects of probucol. In the present study, we compared the effects of probucol (PROB) with that of lovastatin (LOV), a lipid-lowering drug, and trolox (TRO), an antioxidant, on adriamycin (ADR)-induced subchronic in vivo changes in serum free fatty acids (FFA), serum albumin and myocardial reduced (GSH) and oxidized (GSSG) glutathione in rats. ADR caused a significant increase in FFA, decrease in albumin, and an increase in FFA/albumin. PROB and LOV modulated the increases in FFA and FFA/albumin, while TRO was without any effect. ADR reduced myocardial GSH, increased GSSG and decreased GSH/GSSG. Only PROB caused significant improvement in GSH and normalized GSSG levels. It is suggested that these modulatory effects of probucol may also contribute in the beneficial effects of this drug against adriamycin-induced cardiomyopathy and congestive heart failure.

The role of oxidative stress in the genesis of heart disease.

Although researchers in radiation and cancer biology have known about the existence of free radicals and their potential role in pathobiology for several decades, cardiac biologists only began to take notice of these noxious species in the 1970s. Exponential growth of free radical research occurred after the discovery of superoxide dismutase in 1969. This antioxidant enzyme is responsible for the dismutation of superoxide radical--a free radical chain initiator. A fine balance between free radicals and a variety of endogenous antioxidants is believed to exist. Any disturbance in this equilibrium in favour of free radicals causes an increase in oxidative stress and initiates subcellular changes leading to cardiomyopathy and heart failure. Our knowledge about the role of free radicals in the pathogenesis of cardiac dysfunction is fast approaching the point where newer therapies employing antioxidants are in sight.

Effects of afterload-reducing drugs on pathogenesis of antioxidant changes and congestive heart failure in rats.

OBJECTIVES: The present study sought to evaluate the effects of the afterload-reducing drugs captopril and prazosin on changes in antioxidants as well as oxidative stress in relation to hemodynamic function in congestive heart failure (CHF) subsequent to myocardial infarction (MI). BACKGROUND: Afterload reduction therapy has been shown to reduce morbidity and mortality in patients with MI. CHF subsequent to MI in rats is associated with a decrease in myocardial endogenous antioxidants and an increase in oxidative stress. METHODS: The left anterior descending coronary artery in male Sprague-Dawley rats was ligated. Sham and experimental (post-MI [PMI]) animals were assessed for hemodynamic function as well as lung and liver weights at 1, 4 and 16 weeks after operation. At 4 weeks, some rats were also treated with captopril (2 g/liter in drinking water daily) or prazosin (0.2 mg/kg body weight subcutaneously daily) and assessed at 16 weeks. Hearts were isolated to study the activity of superoxide dismutase (SOD), glutathione peroxidase (GSHPx) and catalase as well as for thiobarbituric acid reactive substances (TBARS). RESULTS: CHF at 4 and 16 weeks in the infarcted rats was indicated by an increase in left ventricular end-diastolic pressure and wet/dry weight lung and liver ratios and depressed left ventricular systolic pressure and dyspnea. All these changes were attenuated in both the captopril- and prazosin-treated groups. SOD, GSHPx and catalase activity in the untreated PMI groups was decreased at 4 and 16 weeks. However, treatment with captopril resulted in a significant improvement in SOD, GSHPx and catalase activity in the 16-week PMI group. With prazosin, only SOD activity was improved in the treated 16-week PMI group. Lipid peroxidation as indicated by TBARS was significantly increased in the 16-week PMI group, and both captopril and prazosin modulated this increase. CONCLUSIONS: Occurrence of an antioxidant deficit and an increase in oxidative stress in the myocardium may play a role in the pathogenesis of CHF subsequent to MI. Attenuation of these changes in antioxidant activity with vasodilator (or antioxidant?) therapy mitigates the process of heart failure.

Chronic treatment with propranolol induces antioxidant changes and protects against ischemia-reperfusion injury.

The goal of this study was to examine whether chronic administration of propranolol offers protection against ischemia-reperfusion injury and whether it induces any change in the myocardial endogenous antioxidant enzyme activities and their gene expression. Rats were treated with propranolol (10 mg/kg/day, i.p.) for either 6 or 18 days. Forty-eight h after the last propranolol injection, isolated hearts were subjected to 60 min of global ischemia and 40 min of reperfusion. Resting tension in the control and treated groups after ischemia was 385+/-30 and 150+/-15%; and upon reperfusion was 140+/-11 and 49+/-6%, respectively, as compared to the pre-ischemic values. Recovery of the contractile function in globally ischemic hearts upon reperfusion was about 35% in the treated group as compared to about 16% in the control group at 10 and 20 min. A positive response to catecholamine was observed in hearts from propranolol group (C, 3.41+/-0.36; epi, 6.03+/-0.47 g/g) and was comparable to control hearts (C, 3.55+/-0.31; epi, 6.48+/-0.42 g/g). Myocardial antioxidants, catalase and glutathione peroxidase enzyme activities, in the treated group, prior to ischemia-reperfusion were increased by 67+/-9 and 45+/-11%, respectively, over those in controls. Superoxide dismutase activity did not show any change. The mRNA expression for the three antioxidant enzymes did not change in the hearts of the treated group as compared to control. Lipid peroxidation, both before and after the ischemia-reperfusion episode, was significantly reduced in the propranolol-treated hearts compared to the control group. Hearts studied at the end of reperfusion showed no difference in enzyme activities between treated and control groups. These data show that propranolol treatment of the animals protects against ischemia-reperfusion injury in isolated hearts in the absence of beta-blockade. Increased endogenous antioxidant enzyme activities due to propranolol treatment may have a role in this protection.

Anti-free radical mechanisms in captopril protection against reperfusion injury in isolated rat hearts.

OBJECTIVE: Captopril, an angiotensin-converting enzyme (ACE) inhibitor, is known to modulate ischemia-reperfusion injury in the isolated hearts. This study was designed to examine the involvement of anti-free radical mechanisms in this protection. METHODS: Isolated perfused rat hearts were subjected to 60 mins of global ischemia and 30 mins of reperfusion with or without captopril (100 mumol/L). Myocardial resting tension and contractile force were recorded. At the end of reperfusion, hearts were analyzed for the activities of antioxidant enzymes, superoxide dismutase, glutathione peroxidase and catalase, as well as for the extent of lipid peroxidation. Another potent ACE inhibitor, enalapril (100 mumol/L) was used for comparison. RESULTS: Captopril significantly improved the recovery of contractile function as well as attenuated the rise in resting tension in the ischemic-reperfused hearts as compared to the control. Captopril-exposed ischemic-reperfused hearts showed an increase in the activity of superoxide dismutase with no change in glutathione peroxidase and catalase enzyme activities. Lipid peroxidation at the end of reperfusion was significantly attenuated in the captopril-exposed hearts compared to the control. Enalapril had no protective effect against ischemia-reperfusion induced contractile failure or rise in resting force. CONCLUSIONS: These results suggest that cardioprotection by captopril, against ischemia-reperfusion injury, may involve an anti-free radical mechanism independent of its ACE inhibition property.

Complete stunning in hypertrophied guinea pig heart. Defects in sarcolemmal calcium influx.

Isolated sham control as well as hypertrophied guinea pig hearts were subjected to global ischemia and reperfusion. Developed force declined to zero during 5 min of ischemia without any significant change in resting tension in both sham control and hypertrophied hearts. Upon reperfusion, control hearts showed nearly complete recovery of developed force within 20 min, whereas hypertrophied hearts during this time showed no contractile function, i.e., "a complete stunning" was observed. A continued reperfusion of the stunned hypertrophied hearts ultimately resulted in complete recovery of force within 40-60 min. Data on myocardial cation content showed a relative calcium deficiency in the stunned hearts (3.4 mumol/gm dry wt) as compared to sham control hearts (5.3 mumol/gm dry wt). Stunning could be reversed sooner by isoproterenol (100 microns), and low Na+ (35 and 60 mM) perfusion. Recovery of contractile function by low Na+ was blocked by amiloride (0.17-1.2 mM) in a dose-dependent manner. Perfusion with Bay K8644 (0.1-10 microM) as well as low (0.62 mM) and high (2.5 mM) extracellular calcium concentrations failed to reverse stunning. The pharmacological interventions that were able to reverse the stunning condition also increased the myocardial calcium content. Although the possibilities of a sarcoplasmic reticular dysfunction and/or reduced sensitivity of myofilaments are not excluded, data suggest that a defect in calcium influx across the sarcolemma is an important factor in "complete stunning." It is suggested that this "potential sarcolemmal defect" in the hypertrophied heart, which is unmasked by the ischemic stress, may also represent an early abnormality in the pathogenesis of heart failure.

Probucol treatment reverses antioxidant and functional deficit in diabetic cardiomyopathy.

Earlier we reported that probucol treatment subsequent to the induction of diabetes can prevent diabetes-associated changes in myocardial antioxidants as well as function at 8 weeks. In this study, we examined the efficacy of probucol in the reversal of diabetes induced myocardial changes. Rats were made diabetic with a single injection of streptozotocin (65 mg/kg, i.v.). After 4 weeks of induction of diabetes, a group of animals was treated on alternate days with probucol (10 mg/kg i.p.), a known lipid lowering agent with antioxidant properties. At 8 weeks, there was a significant drop in the left ventricle (LVSP) and aortic systolic pressures (ASP) in the diabetic group. Hearts from these animals showed an increase in the thiobarbituric acid reacting substances (TBARS), indicating increased lipid peroxidation. This was accompanied by a decrease in the myocardial antioxidant enzymes activities, superoxide dismutase (SOD) and glutathione peroxidase (GSHPx). Myocardial catalase activity in the diabetic group was higher. In the diabetic + probucol group both LVSP and ASP showed significant recovery. This was also accompanied by an improvement in SOD and GSHPx activities and there was further increase in the catalase activity. Levels of the TBARS was decreased in this group. These data provide evidence that diabetic cardiomyopathy is associated with an antioxidant deficit which can be reversed with probucol treatment. Improved cardiac function with probucol may be due to the recovery of antioxidants in the heart.